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Comparison of Targeted Next‐Generation Sequencing (NGS) and Real‐Time PCR in the Detection of EGFR , KRAS, and BRAF Mutations on Formalin‐Fixed, Paraffin‐Embedded Tumor Material of Non‐Small Cell Lung Carcinoma—Superiority of NGS
Author(s) -
Tuon Katja,
MäkiNevala Satu,
Sarhadi Virinder Kaur,
Wirtanen Aino,
Rönty Mikko,
Salmenkivi Kaisa,
Andrews Jenny M.,
TelarantaKeerie Aino I.,
Hannula Sari,
Lagström Sonja,
Ellonen Pekka,
Knuuttila Aija,
Knuutila Sakari
Publication year - 2013
Publication title -
genes, chromosomes and cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.754
H-Index - 119
eISSN - 1098-2264
pISSN - 1045-2257
DOI - 10.1002/gcc.22047
Subject(s) - kras , targeted therapy , cancer research , mutation , lung cancer , dna sequencing , digital polymerase chain reaction , biology , medicine , cancer , gene , polymerase chain reaction , oncology , genetics
The development of tyrosine kinase inhibitor treatments has made it important to test cancer patients for clinically significant gene mutations that influence the benefit of treatment. Targeted next‐generation sequencing (NGS) provides a promising method for diagnostic purposes by enabling the simultaneous detection of multiple mutations in various genes in a single test. The aim of our study was to screen EGFR , KRAS , and BRAF mutations by targeted NGS and commonly used real‐time polymerase chain reaction (PCR) methods to evaluate the feasibility of targeted NGS for the detection of the mutations. Furthermore, we aimed to identify potential novel mutations by targeted NGS. We analyzed formalin‐fixed, paraffin‐embedded (FFPE) tumor tissue specimens from 81 non‐small cell lung carcinoma patients. We observed a significant concordance (from 96.3 to 100%) of the EGFR , KRAS , and BRAF mutation detection results between targeted NGS and real‐time PCR. Moreover, targeted NGS revealed seven nonsynonymous single‐nucleotide variations and one insertion‐deletion variation in EGFR not detectable by the real‐time PCR methods. The potential clinical significance of these variants requires elucidation in future studies. Our results support the use of targeted NGS in the screening of EGFR , KRAS , and BRAF mutations in FFPE tissue material. © 2012 Wiley Periodicals, Inc.